Exploring the Association of Surface Plasmon Resonance with Recombinant MHC:Ig Hybrid Protein as a Tool for Detecting T Lymphocytes in Mice Infected with Leishmania (Leishmania) amazonensis.

Lenilton Silva Da Silveira-Júnior,Geraldo Barroso Cavalcanti Júnior,Franklin Souza-Silva,Carlos Roberto Alves,Léa Cysne-Finkelstein,Bernardo Acácio Santini Pereira

doi:10.1155/2017/9089748

Lenilton Silva Da Silveira-Júnior, Geraldo Barroso Cavalcanti Júnior + Show 4 more

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https://doi.org/10.1155/2017/9089748

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Abstract

A surface plasmon resonance- (SPR-) based recognition method applying H-2 Ld:Ig/peptides complexes for ex vivo monitoring cellular immune responses during murine infection with Leishmania (Leishmania) amazonensis is described. Lymphocytes from lesion-draining popliteal lymph nodes were captured on a carboxylated sensor chip surface previously functionalized with H-2 Ld:Ig (DimerX) protein bound to synthetic peptides derived from the COOH-terminal region of cysteine proteinase B of L. (L.) amazonensis. In computational analysis, these peptides presented values of kinetic constants favorable to form complexes with H-2 Ld at neutral pH, with a Gibbs free energy ΔG° < 0. The assayed DimerX:peptide complexes presented the property of attaching to distinct T lymphocytes subsets, obtained from experimentally infected BALB/c mice, in each week of infection, thus indicating a temporal variation in specific T lymphocytes populations, each directed to a different COOH-terminal region-derived peptide. The experimental design proposed herein is an innovative approach for cellular immunology studies of a neglected disease, providing a useful tool for the analysis of specific T lymphocytes subsets.

Highlights

According to the World Health Organization, leishmaniasis is one of the most neglected diseases in the world with extremely limited investments in the development of diagnosis, treatments, or control management
It is estimated that 12 million cases exist worldwide, with 2 million new cases occurring each year: 0.5 million of visceral leishmaniasis (VL) and 1.5 million of cutaneous leishmaniasis (CL) [1, 2]
It was possible to simulate some aspects of the antigen presentation via major histocompatibility complex (MHC) class I to T lymphocytes, once the used fusion protein has characteristics that allow for reproducing in vitro, with reasonable accuracy, the conditions in which epitopes/MHC/T cells receptor interactions occur in vivo [20]

Summary

Introduction

According to the World Health Organization, leishmaniasis is one of the most neglected diseases in the world with extremely limited investments in the development of diagnosis, treatments, or control management (http://www.who.int/ neglected diseases/diseases/en). The disease is endemic in tropical and subtropical regions, covering more than 90 countries and territories, affecting mainly the population of developing countries, where more than 350 million people are at risk. The term leishmaniasis comprises a set of clinical manifestations resulting from infection by the obligate intracellular protozoan parasites from the genus Leishmania. One species belonging to this genus is the Leishmania (Leishmania) amazonensis, which is an important etiologic agent of cutaneous leishmaniasis in humans, perhaps most importantly, with varied clinical manifestations in their infections [3, 4]

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